1.The changes of mRNA expression of telomerase subunits induced by Ara-C in HL-60 cells
Ji-Hua ZHONG ; Fang-Yuan CHEN ; Hong-Hui WANG ; Hai-Rong HANG ; Ren-Rong OUYANG ;
China Oncology 2006;0(09):-
Background and purpose:Ara-C is one of the most effective and common agents in the treatment of acute nonlyphocytic leukemia. Telomerase is a unique complex of ribonucleoprotein. It plays an important role in the pathogenesis and development of cancer. In this study, we investigate the changes of mRNA expression of telomerase subunits in HL-60 cells induced by Ara-c and try to come up with a theory that could help to assess the efficacy of Ara-C. Methods:The combinations of various Ara-C concentration and the incubation time were used to treat HL-60. The ratios of apoptotic cell to necrosis cell were determined by flow cytometry and the expressions of telomerase subunits mRNA were evaluated by RT-PCR.Results:① There was no influence on transcription of telomerase subunits gene after HL-60 cells was cultured with 0~0.2ug/ml Ara-C for 12 hours;② 2ug/ml and 10ug/ml of Ara-C could down regulate the expression of hTERT from 0.80+0.07 to 0.50+0.04 and 0.39+0.03, not hTR and hTP1;③ with longer incubation with 10ug/ml of Ara-C, the percentage of apoptosis could be increased. The maximal induction of apoptosis (18.16+4.25%) could be reached at 12hrs treatment of Ara-C, then gradually decreased later on. The rate of necrosis increased with time, the maximal percentage(57.94+12.03%) of necrosis was observed at 48hrs of incubation time with drug. The mRNA level of hTERT gene also decreased along with the cultured time , the lowest value (0.18+0.03) has been documented at 48hrs time point, but not hTR、TP1.Conclusions:① Ara-C could down-regulate the expression of hTERT mRNA in a dose-and time-dependent manner, but not hTR、hTP1;② There might be no relationship between the percentage of apoptosis induced by Ara-C apoptosis and the expression of telomerase hTERT gene mRNA, but a close relationship between necrosis and the expression of hTERT mRNA has been found.
2.Biological activity of survivin antisense oligonucleotide labeled with quantum dots or green fluorescein: a comparative study.
Hang YAO ; Zong-hai HUANG ; Zhou LI ; Rong HE ; Feng GAO ; Da-xiang CUI
Journal of Southern Medical University 2007;27(5):663-666
OBJECTIVETo compare the durability of quantum dots with that of green fluorescein for labeling survivin antisense oligonucleotide (ASODN) and investigate the difference in growth and apoptosis of cells transfected with the labeled survivin ASODN.
METHODSSurvivin ASODN labeled with quantum dots or green fluorescein was transfected into MCF-7 cells via Lipolifectmain(TM2000). The proliferation of MCF-7 cells was assessed with MTT assay, survivin mRNA expression determined by RT-PCR and its protein expression measured by Western blot analysis. The apoptosis rate of the transfected cells was estimated by flow cytometry, and the fluorescence distribution in the cells observed under fluorescent inverted microscope.
RESULTSThe mRNA and protein expressions of survivin were significantly decreased in the MCF-7 cells after cell transfection with survivin ASODN labeled with quantum dots or green fluorescein, and no significant difference was noted between the two labeling methods (P>0.05). Nor did survivin ASODN transfection with different labeling methods produced significant difference in cell proliferation and apoptotic rate (P>0.05). For green fiuorescein labeling, the fluorescence disappeared 4 days after transfection, whereas the fluorescence sustained for 1 week for quantum dots labeling.
CONCLUSIONSurvivin ASODNs labeled with quantum dots and green fiuorescein do not significantly differ in survivin expression or the transfected cell proliferation and apoptosis rate, but quantum dot labeling can be more stable with longer maintcnance of the labeling.
Apoptosis ; Blotting, Western ; Cell Line, Tumor ; Cell Proliferation ; Flow Cytometry ; Fluorescein ; chemistry ; Gene Expression ; Humans ; Inhibitor of Apoptosis Proteins ; Microscopy, Fluorescence ; Microtubule-Associated Proteins ; genetics ; metabolism ; Oligonucleotides, Antisense ; chemistry ; genetics ; Quantum Dots ; Reverse Transcriptase Polymerase Chain Reaction ; Staining and Labeling ; methods ; Transfection
3.Evaluating on recognition impairment after traumatic brain injury with WCST.
Rong-Hua HANG ; Ya-Jun XU ; Hai-Feng XIE ; Xu-Yang ZHU
Journal of Forensic Medicine 2011;27(5):346-349
OBJECTIVE:
To explore the value of Wisconsin Card Sorting Test (WCST) in evaluating the recognition impairment after traumatic brain injury (TB1).
METHODS:
WCST and Wechsler Adult Intelligence Scale Revised in China (WAIS-RC) were adopted to assess the cognitive function in 186 patients with brain injury (experimental group) and 180 healthy volunteers (control group), respectively. The cognition between the experimental group and the control group was compared. The correlation between WCST and IQ was analyzed. The correlation between age, education level, the severity of TBI and the executive function were also analyzed.
RESULTS:
(1) The performances of WAIS-RC and WCST in the experimental group were significantly worse than that of the control group. (2) There were significantly negative correlation between the performance of total errors, percentage of preservative errors, percentage of random errors, response number on first category and IQ (P < 0.05). While there were significantly positive correlation between the categories completed, percentage of conceptual level and IQ (P < 0.05). (3) There was significantly negative correlation between percentage of conceptual level and the severity of TBI. While there were significantly positive correlation between percentage of preservative errors, percentage of random errors, response number on first category and the severity of TBI.
CONCLUSION
WCST could be an important method in evaluating cognition of patients with brain injury.
Adolescent
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Adult
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Aged
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Brain/pathology*
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Brain Injuries/psychology*
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Case-Control Studies
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Cognition Disorders/etiology*
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Executive Function/physiology*
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Female
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Humans
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Male
;
Middle Aged
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Multivariate Analysis
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Neuropsychological Tests
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Predictive Value of Tests
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Psychiatric Status Rating Scales
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Psychometrics
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Reproducibility of Results
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Severity of Illness Index
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Task Performance and Analysis
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Wechsler Scales
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Young Adult
4.Estimation and prediction of the epidemic situation of acquired immunodeficiency syndrome in Jiangsu province.
Na LEI ; Zhi-hang PENG ; Xi-ping HUAN ; Xin CHEN ; Rong-bin YU ; Hai-tao YANG
Chinese Journal of Preventive Medicine 2010;44(11):1012-1017
OBJECTIVEThe aim of this study was to estimate and predict the AIDS epidemic situation in Jiangsu province by using a software named Estimation and Projection Package (EPP) and then to provide accurate information for making control plans and conducting interventions.
METHODSDate were obtained from the comprehensively monitored sentinels of AIDS and venereal diseases (Data were from 28 national monitored stations and 52 provincial stations. From 2003 to 2009, a total of 10 730 000 people had been monitored) as well as project survey and laboratory tests. EPP epidemic model was employed to analyze the prevalence and to predict future epidemic situations.
RESULTSA total of 830 000 and 1 020 000 people were monitored in 2003 and 2004 respectively, and 8 880 000 more people were monitored in the following 5 years. By the end of 2009, a total of 4103 HIV infection cases had been reported, among which 918 people were diagnosed with AIDS and 432 died. Since 2003, the cases infected through IDU had decreased from 66.84% (262/392) to 16.40% (142/868). However, the cases infected through sex contact had increased from 21.68% (85/392) to 77.40% (672/868). Among these cases, homosexual transmission and heterosexual transmission accounted for 39.10% (339/868) and 38.30% (333/868) respectively. It was estimated that there would be 14 290 HIV/AIDS cases in 2011 which is 18.10% (2191/12 099) higher than that in 2009. The estimated HIV infection rate would be 0.02%.
CONCLUSIONEPP prediction showed the AIDS epidemic situation in Jiangsu province had an ascendant trend and the AIDS epidemic situation had demonstrated rapid growth. Therefore, it is necessary to take effective preventive measures to control the spread of AIDS.
Acquired Immunodeficiency Syndrome ; epidemiology ; prevention & control ; China ; epidemiology ; Epidemics ; statistics & numerical data ; Forecasting ; HIV Infections ; epidemiology ; prevention & control ; Humans ; Prevalence ; Sentinel Surveillance ; Software ; Statistics as Topic
5.Specific killing effects of combination of recombinant adenovirus containing double suicide gene driven by KDR promoter and survivin antisense oligonucleotide on colorectal cancer cells and vascular endothelial cells.
Hang YAO ; Zong-hai HUANG ; Zhou LI ; Guo-qiang SU ; Rong HE ; Feng GAO ; Da-xiang CUI
Chinese Journal of Gastrointestinal Surgery 2008;11(1):61-66
OBJECTIVETo evaluate the specific killing effects of combination of recombinant adenovirus mediated double suicide gene driven by KDR promoter and survivin antisense oligonucleotide(ASODN) on colorectal cancer cells and vascular endothelial cells.
METHODSThe 293 packaging cells were transfected with the plasmids of pAdEasy-CDglyTK and the recombinant adenovirus were generated. The KDR expressive cells of SW620, ECV304 were infected with adenovirus, meanwhile survivinASODN was transferred into the same cells. The infection rate of adenovirus and transfection efficiency of survivinASODN were observed and the expression of CDglyTK was detected by RT-PCR. The expression of survivin was measured by Western blot. The killing effects and bystander effects on SW620, ECV304 were examined through MTT method.
RESULTSThe cells which were infected with the adenovirus mediated double suicide gene could be transfected with the survivin ASODN and the infection rate was not affected as well as the transfection efficiency. The high expression of CDglyTK gene was found in SW620, ECV304 cells infected with recombinant adenovirus and survivin ASODN decreased the survivin protein level. The survival rate of gene therapy group was significantly lower than that of negative group. The combination of survivin ASODN and AdKDR-CDglyTK gene therapy showed significantly lower survival rate of SW620 and ECV304 cells as compared with the AdKDR-CDglyTK or survivin ASODN used alone (P<0.05). The survival rate was slightly lower in GCV 100 microg/ml, 5-FC 2000 microg/ml than that AdKDR-CDglyTK used alone (P>0.05). The combined therapy of AdKDR-CDglyTK and survivin ASODN showed synergistic killing efficacy and more significant bystander effects.
CONCLUSIONThe combined gene therapy of AdKDR-CDglyTK system and survivin ASODN has stronger specific killing effects on colorectal cancer cells and vein endothelial cells.
Adenoviridae ; genetics ; Cell Line, Tumor ; Colorectal Neoplasms ; genetics ; metabolism ; Endothelial Cells ; metabolism ; Genes, Transgenic, Suicide ; genetics ; Humans ; Inhibitor of Apoptosis Proteins ; Microtubule-Associated Proteins ; genetics ; Oligonucleotides, Antisense ; genetics ; Receptors, Vascular Endothelial Growth Factor ; genetics ; Transcription Initiation Site
6.Specific killing effects of combination of double suicide gene and survivin antisense oligonucleotide on breast tumor cells and vein endothelial cells.
Hang YAO ; Zong-Hai HUANG ; Zhou LI ; Guo-Qiang SU ; Rong HE ; Feng GAO ; Da-Xiang CUI
Chinese Journal of Surgery 2007;45(7):476-479
OBJECTIVETo evaluate the selectively killing effects of combination of adenovirus mediated double suicide gene driven by kinase-domain insert containing receptor (KDR) promoter and survivin antisense oligonucleotide on breast tumor cells and vein endothelial cells.
METHODSHuman embryonal kidney cells 293 were transfected with the plasmids of pAdEasy-KDR-CDglyTK and the adenovirus was generated. The KDR expressive cells of MCF-7, ECV304 were infected by adenovirus and survivin ASODN was transferred into the same cells meanwhile. The infection rates of adenovirus and transfection efficiency of survivin ASODN were observed and the expression of CDglyTK was detected by RT-PCR. The expression of survivin was measured by Western blot. The killing effects and bystander effects on cells were assessed by MTT assay.
RESULTSThe cells infected by the adenovirus mediated double suicide gene could be transfected with the survivin ASODN and the infection rate was not affected as well as the transfection rate. The high expression of CDglyTK gene was found in the two kinds of cells and survivin ASODN decreased the survivin protein level. When survivin ASODN was transferred into MCF-7, ECV304 cells, the survival rates were 56.4% +/- 3.8% and 55.9% +/- 3.6% respectively. The combination of survivin ASODN and AdKDR-CDglyTK gene therapy showed significantly lower survival rate comparing with using each treatment alone (P < 0.05) and the survival rate decreased gradually with the increasing of the concentration of GCV and 5-FC. But the survival rate for combined gene therapy group was slightly lower in GCV 100 microg/ml, 5-FC 2000 microg/ml than that of AdKDR-CDglyTK group (P > 0.05). The combination of survivin ASODN and AdKDR-CDglyTK therapy showed synergistic killing efficacy and more significant bystander effects.
CONCLUSIONThe combined therapy with AdKDR-CDglyTK system and survivin ASODN shows obvious killing effects on breast tumor cells and vein endothelial cells.
Adenoviridae ; genetics ; Breast Neoplasms ; genetics ; metabolism ; pathology ; Cell Line ; Cell Line, Tumor ; Cell Survival ; Endothelial Cells ; metabolism ; pathology ; Female ; Genes, Transgenic, Suicide ; genetics ; Genetic Therapy ; methods ; Genetic Vectors ; Humans ; Inhibitor of Apoptosis Proteins ; Microtubule-Associated Proteins ; genetics ; Neoplasm Proteins ; genetics ; Oligonucleotides, Antisense ; genetics ; Plasmids ; Promoter Regions, Genetic ; Receptors, Vascular Endothelial Growth Factor ; genetics ; Transfection
7. Mechanism of 1,2-dichloroethane-induced toxicity in SH-SY5Y Cells
Si LI ; Hong-Ling LI ; Na ZHAO ; Xiang-Rong SONG ; Hao-Zhong LIU ; Jia-Bin CHEN ; Long-Yuan JIANG ; Yong-Shun HANG ; Hai-Lan WANG
China Occupational Medicine 2016;43(06):652-661
OBJECTIVE: To establish the cell model of human neuroblastoma cell( SH-SY5Y cell) exposed to1,2-dichloroethane( 1,2-DCE) in vitro and to explore the mechanism of 1,2-DCE-induced toxicity in SH-SY5Y cells.METHODS: SH-SY5Y cells were collected in their logarithmic growth phase and cultured in complete medium that had final concentrations of 1,2-DCE in 0,10,20,30,40,50,60,70 and 80 mmol / L for 24 hours. Cell morphology was observed and cell survival rate was examined by CCK-8 assay. Using chemical colorimetric method, the activity of lactic dehydrogenase( LDH) in the cell culture supernatant,and the intracellular level of malondialdehyde( MDA),the intracellular activities of superoxide dismutase( SOD) and adenosine triphosphate( ATP) enzymes were detected. RESULTS: With the increasing exposure concentrations of 1,2-DCE,the cell density of SH-SY5Y cells gradually decreased,the synapse became shorter,the membrane ruptured,cytoplasm condensed and cytoplasmic contents overflowed increased.With the increasing concentration of 1,2-DCE,the cell survival rate decreased( P < 0. 01),the activity of LDH in the cell culture supernatant increased( P < 0. 01). These changes had a dose-effect correlation. Intracellular MDA level,and activities of SOD,Na~+-K~+-ATP enzyme,Ca~(2+)-Mg~(2+)-ATP enzyme and total ATP enzyme increased at first and then decreased. The activity of LDH in the cell culture supernatant and cell survival rate was negatively correlated( the correlation coefficient is- 0. 907,P < 0. 01). CONCLUSION: 1,2-DCE could inhibit the proliferation of SH-SY5Y cells.The mechanism may be related to the permeability change of cell membrane,cellular damage from excessive free radicals,the decrease of free radical scavenging capacity,ATP enzyme activity and calcium overloading. SH-SY5Y cells can be used as a common cell line for 1,2-DCE cytotoxicity analysis.
8.Study on the spatial distribution of AIDS based on geographic information system in Jiangsu province
Zhi-Hang PENG ; Hai-Tao YANG ; Yue-Jia CHENG ; Lu WANG ; Rong-Bin YU ; Xi-Ping HUAN ; Hong-Bing SHEN ; Feng CHEN ; Ning WANG
Chinese Journal of Epidemiology 2011;32(1):42-46
Objective To analyze the spatial distribution of AIDS in every city of Jiangsu province, trying to describe the geographic characteristics of AIDS and to develop a prediction model.Methods Numbers of patients in Jiangsu province were collected, to establish the database for the geographic information system, then setting up a 'risk map' of the disease. Spatial, autocorrelation.Linear spatial analyses were used to study the patients' numbers. Results (1)Results from the autocorrelation analysis showed that the distribution of AIDS was clustered at some places and was at random on the whole. The results also indicated that the distribution of AIDS in Nanjing was of negative correlation, while that in Suzhou and Wuxi were of positive correlation but in Tongshan, Wujiang, Pukou, Nanjing, Lishui, Wuxi and Suzhou showed seven locations of clusters with disease was more severe in the southern than in the northern parts of Jiangsu province. (3)Ordinary Least Squares method was finally used in the linear spatial regression and the results were: t=-1.045 103 (P=0.299 904); t=-1.443 668 (P=0.153 714) respectively. Conclusion According to the feature spatial distribution of the disease, effective measures should be taken to prevent and to keep the prevalence of AIDS under control.
9. Anti-lung Cancer Mechanism of Natural Quinonoids
Hai-rong ZHONG ; Chun-pu ZOU ; Zi-hang XU
Chinese Journal of Experimental Traditional Medical Formulae 2019;25(14):32-44
Lung cancer is a common malignancy with a high morbidity and high mortality. Although chemoradiotherapy is still the main therapy, the side effects caused by radiotherapy and chemotherapy cannot be ignored. There is an urgent need to find a targeted anti-cancer compound with a low toxicity and strong effect. Therefore, a large number of natural product-derived compounds have received increasing attention for their anti-cancer effect. They are diverse in structure, rich in variety but simpler in composition than natural plants, with a clear efficacy. This facilitates research, and the enriched ingredients have stronger pharmacological effect and clinical efficacy than natural plants. Quinonoids are natural bioactive molecules that widely exist in more than 100 species of higher and lower plants, including naphthoquinones, phenanthrenequinones, benzoquinones and pyrenes. Most of them have anti-tumor effect. They are applied and studied in many cancers, especially lung cancer; and the mechanism of action involves multiple pathways, which play a role in promoting apoptosis, inhibiting proliferation, inducing autophagy, inhibiting angiogenesis and cell invasion and migration. When they are used in combination with other drugs, there is also an anti-tumor effect. With the development of researches, many synthetic quinonoid derivatives with similar structures also increase the diversity of anti-cancer steroids, and quinonoids have broad application prospects in lung cancer. On the basis of a large amount of literature, this article reviews current studies on the molecular mechanisms of antitumor activity of quinonoids, summarizes the common research targets with different effect, and analyzes the relationship between the pharmacological effects and anti-lung cancer effects of quinonoids, and proposes the direction of further research, so as to provide the reference for the follow-up research and development of other quinonoids.
10.Design and biomechanical study of a novel double-leaf proximal humeral locking plate
Lang-qing ZENG ; Lu-lu ZENG ; Yun-feng CHEN ; Yan-jie LIU ; Hui YANG ; Zhi-rong LIN ; Wen HANG ; Hai-feng WEI
Journal of Medical Biomechanics 2017;32(5):E407-E414
Objective To design a novel double-leaf proximal humeral locking plate for fixing greater and lesser tuberosities in complex proximal humeral fractures, and evaluate its fixing stability by biomechanical tests. Methods Twelve fresh-frozen humerus specimens with intact rotator cuff were randomly divided into two groups (Group A and Group B) to establish the same greater and lesser tuberosities fracture models. Specimens in Group A were fixed with the double-leaf proximal humeral locking plate, while specimens in Group B were fixed with the proximal humeral internal locking system (PHILOS) and tension band suture, and a 3.5-mm cannulated screw was added to stabilize the lesser tuberosity. The tensile test on subscapularis, infraspinatus and teres, supraspinatus as well as the load-to-failure test on greater and lesser tuberosities were performed on specimens in two groups. Results For subscapularis tensile tests, displacements under 150 N tensile stretch and after fatigue test in Group A were both significantly smaller than those in Group B (P<0.05). For infraspinatus and teres tensile tests, there were no statistical differences between Group A and B in displacements under 150 N tensile stretch and after fatigue test (P>0.05). For supraspinatus tensile tests, there were no statistical differences between Group A and B in displacements under 90 N tensile stretch and after fatigue test (P>0.05). For load-to-failure tests on lesser tuberosity, the failure load in Group A was significantly greater than that in Group B (P<0.05), and the failure displacement in Group A was significantly smaller than that in Group B (P<0.05). For load-to-failure tests on greater tuberosity, there were no statistical differences between Group A and B in both the failure load and failure displacement (P>0.05). Conclusions Compared with the ordinary tension band suture plus cannulated screw for fixing lesser tuberosity, the novel double-leaf proximal humeral locking plate shows more obvious biomechanical stability, with the advantage of simultaneously fixing greater and lesser tuberosities. The research findings provide a new choice for the clinical treatment of complex proximal humeral fractures.